The NADPH oxidase DUOX1 is prominently expressed within the respiratory epithelium and has been invoked in innate airway epithelial responses to injury or infectious stimuli, by promoting H2O2-dependent signaling pathways that mediate wound responses and production of various inflammatory mediators or mucin genes. In studies during the previous funding cycle, we have identified oxidative mechanisms by which DUOX1 activation promotes the activation of epidermal growth factor receptor (EGFR) as a critical event in wound responses and inflammatory mediator production, and observed increased DUOX1 expression in models of allergic asthma in association with increased EGFR activation and mucus metaplasia. The respiratory epithelium is critical in coordinating innate and adaptive immune responses to airborne allergens, by production of epithelial cytokines such as interleukin-33 (IL-33). IL-33 was recently identified as a critical mediator in animal models of allergic inflammation, and a major susceptibility gene for asthma. IL-33 is normally present within the nuclei of pulmonary epithelial cells, and is rapidly secreted in response to various stimuli and allergens to function as an alarmin that stimulates ST2 receptors on target cells to induce innate TH2 cell-dominant immune responses. The cellular mechanisms involved in regulating IL-33 secretion are still poorly understood, however, and we now present preliminary evidence demonstrating the importance of DUOX1-dependent production of H2O2 as a critical mediator in epithelial IL-33 secretion in response to two common allergens, Alternaria alternata and Dermatophagoides pteronyssinus (house dust mite). Our studies further indicate the importance of protease-activated receptors (PAR) and transient receptor potential (TRP) channels in proximal responses to these allergens leading to DUOX1 activation and IL-33 secretion. In addition, we have preliminary evidence demonstrating H2O2-mediated EGFR activation as a critical mechanism by which DUOX1 activation promotes nuclear export and/or secretion of IL-33. The main objectives of this renewal application are to establish the common role of DUOX1 activation in mediating IL-33 secretion and subsequent allergic inflammation by various protease allergens (Aim 1), to clarify the downstream redox mechanisms by which DUOX1 activation promotes IL-33 nuclear export and secretion, focusing on the role of EGFR activation (Aim 2), and to expand our findings in studies with human subjects with allergic asthma by determining associations between DUOX1 expression and activation in nasal epithelial allergen responses in these patients (Aim 3). Successful accomplishment of these studies would identify DUOX1 as a novel and attractive therapeutic target in the management of asthma and/or its exacerbations.